Microbiome based agents and uses thereof

Abstract

The subject disclosure relates to microbiome-based agents, compositions and methods for treatment or prevention of pain, osteoarthritis, and other complications in a subject (e.g., a subject with a sickle cell disease). In particular, the present disclosure relates to compositions comprising probiotic microorganisms and uses of such compositions for treating, preventing or reducing pain and osteoarthritis in a subject.

Description

Attorney Docket No.: 45301.601MICROBIOME BASED AGENTS AND USES THEREOFRELATED APPLICATION INFORMATION

[0001] This application claims priority to U.S. Application No. 63 / 684,914 filed on August 20, 2024, the contents of which is herein incorporated by reference.FIELD OF THE DISCLOSURE

[0002] The subject disclosure relates to microbiome-based agents, compositions, and methods for treatment or prevention of pain, osteoarthritis, and other complications in a subject (e.g., a subject with a sickle cell disease). In particular, the present disclosure relates to compositions comprising probiotic microorganisms and uses of such compositions for treating, preventing or reducing pain and osteoarthritis in a subject.BACKGROUND

[0003] Sickle cell disease (SCD) is the most common genetic blood disorder worldwide that impacts millions of individuals. Blockade of capillaries with sickle red blood cells induces vasoocclusive episodes and inflammation, which causes severe pain and organ damage and multiple clinical complications, including osteoarthritis (OA). Degenerative osteoarthropathy is a form of OA prevalent in individuals with SCD and is associated with significant morbidity, with no effective therapy.

[0004] Therefore, the development of therapeutic and preventative approaches to modulate pain, osteoarthritis, and other complications in SCD is needed.SUMMARY

[0005] In an aspect, disclosed is an agent for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject (e.g., a subject with a sickle cell disease), the agent includes at least one microbiome such as a probiotic.

[0006] In an aspect, disclosed is a composition for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject with a sickle cell disease, the composition including a therapeutically effective amount of the agent including a microbiome.Attorney Docket No.: 45301.601

[0007] In an aspect, disclosed is a method for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject with a sickle cell disease, the method including providing and / or administering the subject an agent, a composition, or a combination.

[0008] In particular, provided herein is a method for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising: providing and / or administering a therapeutically effective amount of a composition comprisingLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus to a subject in need thereof under conditions such that pain and / or symptoms of OA are prevented, treated, or reduced in the subject. In certain aspects, the subject has sickle cell disease. In some embodiments, the pain is hyperalgesia and / or hypersensitivity to cold. In some embodiments, the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular bone.

[0009] In some embodiments, the composition is a pharmaceutical composition, for example comprising one or more pharmaceutically acceptable carriers. In one non-limiting example, the carrier is a starch. In certain aspects, the Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus are lyophilized. The present disclosure is not limited to a particular formulation of the composition. Examples include but are not limited to, a powder, capsule, liquid, or paste.

[0010] While not limited to a particular delivery format, in certain aspects, the administering is oral administration. In some embodiments, the composition is provided as a unit dosage.Examples of suitable unit dosages include but are not limited to, a total of about 200 billion to 500 billion colony forming units (CFUs) per dose (e.g., about 200 billion, 225 billion, 250 billion, 300 billion, 350 billion, 400 billion, 450 billion, or 500 billion CFUs per dose). In some embodiments, 1 to 3 doses are administered per day (e.g., for a time period of 1 week to one year to indefinitely).

[0011] Also provided is a composition for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising Lactobacillus casei subsp. paracasei,Attorney Docket No.: 45301.601Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus.

[0012] Further provided is the use of a composition comprisingLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject.

[0013] These and other aspects of the present invention are described in more detail below.

[0014] DESCRIPTION OF THE FIGURES

[0015] FIG. 1 shows that VISBIOME probiotic treatment partially rescues pain behaviors in SCD mice. Pain behaviors analyzed at 5 weeks post treatment for (A) deep tissue hyperalgesia, (B) mechanical hyperalgesia to von Frey (1.0g fiber), (C) thermal sensitivity to cold (C), (D) thermal sensitivity to hot. Data are presented as mean ± SEM. n=4-6 mice / group. *P,0.05 by ANOVA. PWF, paw withdrawal frequency; PWL, paw withdrawal latency.

[0016] FIG. 2 shows that VISBIOME probiotic treatment partially rescues osteoarthritis (OA) phenotype in SCD mice. (A) Representative x-ray images of excised knee joint showed the flattened tibia plateau and uneven joint surfaces in the SCD-Veh mice (arrow) were partially rescued with VISBIOME probiotic (B) Representative microCT images of excised knee joint show erosion of articular surface in the SCD-Veh group (arrow) was partially rescued by VISBIOME probiotic treatment.

[0017] FIG. 3 shows that VISBIOME probiotic treatment partially rescues femoral epiphysis structure parameters in SCD mice. (A) Representative microCT images of excised knee joint showed thinning of trabecular in SCD-Veh group (arrow), which was partially rescued by VISBIOME probiotic. MicroCT analysis of parameters of femoral epiphysis (B) BV / TV, (C) Tb.N, (D) Tb.Th, and (E) TMD. Data are presented as mean ± SEM. n=4-6 mice / group. *P,0.05 by ANOVA.DETAILED DESCRIPTION

[0018] Before the disclosed processes and materials are described, it is to be understood that the aspects described herein are not limited to specific embodiments, or examples, and as suchAttorney Docket No.: 45301.601 can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and, unless specifically defined herein, is not intended to be limiting.

[0019] Recent studies showed gut microbiota dysbiosis in humanized Townes Sickle cell disease (SCD) mice displayed comparable dysbiosis to that seen in humans with SCD and altered gut microbiota play a role in mediating pathogenetic phenotypes of SCD subjects.

[0020] Experiments described herein determined that probiotics, such as commercially available probiotics, for example, VISBIOME probiotic, can rescue pain or osteoarthritis observed in SCD mice.

[0021] Accordingly, described herein are agents, compositions, and methods to provide treatment options for pain, osteoarthritis, and other complications in a subject (e.g., a subject with a sickle cell disease). The present invention relates to agents, compositions, and methods useful for dietary augmentation, to provide a probiotic regimen, and / or to provide therapeutic intervention for the treatment, prevention, amelioration and / or regulation of disease states and / or other adverse physiological conditions, such as pain and osteoarthritis in a subject with a sickle cell disease.

[0022] In an aspect, disclosed is an agent for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject with a sickle cell disease, the agent includes at least one microbiome (as used herein a microbiome is the community of microorganisms such as bacteria, fungi, and viruses that can usually be found living together in any given habitat), such as a probiotic (as used herein a probiotic is a live bacteria and yeasts that have beneficial effects on human body), for example, any suitable commercially available probiotics such as VISBIOME probiotic (includes 8 bacteria species: paracasei, plantarum, acidophilis, delbrueckii subspecies bulgaricus, longum, infantis, breve, thermophilus), its pharmaceutically acceptable salts or derivatives, or a combination thereof.

[0023] In particular, provided herein is a composition for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprisingLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus, or any combinations thereof. In an embodiment, the composition comprises Lactobacillus casei subsp. paracasei, LactobacillusAttorney Docket No.: 45301.601 plantation, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus.

[0024] In some embodiments, the pain is hyperalgesia and / or hypersensitivity to cold. In some embodiments, the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular.

[0025] In an embodiment, other complications include bone loss, such as reduced femoral bone mineral density (BMD). In an embodiment, the agent includes any suitable microbiome, for example, at least one probiotic. In an embodiment, the microbiome is a genetically engineered microbiome. In an embodiment, the agent includes at least one probiotic, its pharmaceutically acceptable salts or derivatives, or a combination thereof. In an embodiment, the probiotic is a commercially available probiotic, for example, VISBIOME probiotic, its pharmaceutically acceptable salts or derivatives, or a combination thereof. In an embodiment, the agent includes VISBIOME probiotic, its pharmaceutically acceptable salts, its derivatives, or a combination thereof. In an embodiment, the subject is a mammal. In an embodiment, the subject is human.

[0026] While not wishing to be bound by this theory, it is believed that the agent acts on inflammatory cell, chondrocyte, osteoblast / osteoclasts and affects their proliferation, differentiation, or function. In an embodiment, the agent is a high-potency probiotic medical food containing at least 8 strains of live bacteria (for example, paracasei, plantarum, acidophilis, delbrueckii subspecies bulgaricus, longum, infantis, breve, thermophilus) scientifically formulated to promote gut health and plays a role in the natural pain and osteoarticular health. In an embodiment, the agent is a high-potency probiotic medical food containing at least 7 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least 6 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least 5 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least 4 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least 3 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least 2 strains of live bacteria. In an embodiment, the agent is a high-potency probiotic medical food containing at least one strain of live bacteria. Any suitable strain of bacteria can be used, some nonlimiting examples are, paracasei, plantarum, acidophilis, delbrueckii subspecies bulgaricus, longum,Attorney Docket No.: 45301.601 infantis, breve, thermophilus, or a combination thereof. The bacterial composition of the present invention, in various embodiments thereof, may comprise, consist of, or consist essentially of the combination of these types of bacteria, and a physiologically compatible carrier medium for such bacteria. In an embodiment, the agent is a probiotic supplement useful in a dietary regimen, and / or to provide therapeutic intervention for the treatment, prevention, amelioration and / or regulation of a pain and osteoarthritis in a subject with a sickle cell disease. In an embodiment, the bacterial species may be in a dried form, e.g., lyophilized or sporolated form, in a suitable carrier medium, e.g., firucto-oligo-saccharide (FOS) medium, or other soluble fiber, sugar, nutrient or base material for the composition, with which the bacterial species can be presented in an orally administrable form.

[0027] In an aspect, disclosed is a composition for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject with a sickle cell disease, the composition including a therapeutically effective amount of the agent including a microbiome as shown and described herein. In an embodiment, the composition further includes a carrier that includes at least one pharmaceutically acceptable excipient. In an embodiment, the composition includes the agent including any suitable microbiome, for example, at least one probiotic. In an embodiment, the composition includes the microbiome, that is a genetically engineered microbiome. In an embodiment, the composition includes the agent including at least one probiotic, its pharmaceutically acceptable salts or derivatives, or a combination thereof. In an embodiment, the composition includes the probiotic that is a commercially available probiotic, for example, VISBIOME probiotic, its pharmaceutically acceptable salts or derivatives, or a combination thereof. In an embodiment, the composition includes the agent including VISBIOME probiotic, its pharmaceutically acceptable salts, its derivatives, or a combination thereof. In an embodiment, the subject is human.

[0028] In an embodiment, the composition includes a carrier medium. In an embodiment, the carrier medium, when present, can be blended with the bacterial species in any suitable amounts, such as an amount of from about 5% to 95% by weight of carrier medium, based on the total weight of the bacterial species and the carrier medium. In certain embodiments, the amount of carrier medium may be in a range having a lower limit of any of about 5%, 10%, 12%, 15%, 20%, 25%, 28%, 30%, 40%, 50%, 60%, 70% or 75%, and an upper limit, higher than the lower limit, of any of 20%, 22%, 25%, 28%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, andAttorney Docket No.: 45301.601 about 95%. In an embodiment, the subject is a mammal. The amount of carrier medium in a specific embodiment may be determined based on considerations of the specific dose form, relative amounts of the three bacterial species, the total weight of the composition including the carrier medium and the bacterial species, and the physical and chemical properties of the carrier medium, and other factors, as known to those of ordinary skill in the probiotic formulation art.

[0029] The compositions of the invention, including the bacterial species and optionally a suitable carrier medium, can be provided in any suitable dose form for administration. The dose form is preferably a pill, powder, capsule, sachet, or the like, formulated for oral administration. In an embodiment, the dose form is a capsule containing the composition as disclosed herein. The capsule for such dose form can be of any suitable type, e.g., a gelatin capsule of a conventional variety. Preferably, such capsule is devoid of any enteric coating. Preferably the bacterial species are non-microencapsulated in dose form.

[0030] The compositions may additionally and optionally include any suitable adjuvants, excipients, additives, carriers, additional therapeutic agents, bioavailability enhancers, side-effect suppressing components, diluents, buffers, flavoring agents, binders, preservatives or other ingredients that do not preclude the efficacy of the composition. In an embodiment, the microbiome including suitable bacteria may include from about 50% to about 90% by weight of the composition, based on the total weight of the composition including a carrier medium, most preferably from about 60% to about 80% by weight of the composition. It is generally preferred that the composition contain only the bacteria and a carrier medium, in a gelatin capsule that is devoid of any enteric coating, with the bacteria being in a dried form such as powder or granules, and the carrier medium likewise being in a dry powder or particulate form. The compositions of the present invention are preferably formulated for oral administration. Other routes of administration can be employed, however, including, but not limited to, subcutaneous, intramuscular, intradermal, transdermal, intraocular, intraperitoneal, mucosal, vaginal, rectal, and intravenous. For example, a dose of the composition of the present invention can be presented in a gelatin capsule and inserted vaginally to treat vaginal yeast infections. The bacterial species therefore are present in the dose form as live bacteria, whether in dried, lyophilized, or sporolated form.

[0031] In an embodiment, the therapeutic composition of the invention may comprise at least 40 million CFUs of each of bacteria, as contained in capsules containing from about 25 to 200Attorney Docket No.: 45301.601 milligrams of the bacteria or the mixture of bacteria with one or more excipients, such as about 500-1,000 milligrams of fructo-oligosaccharides, in a unitary dose form tablet or capsule.

[0032] In an embodiment, the agent, the composition, or a combination thereof that can be used (a therapeutic amount) is about 25 million to about 2 billion live colony forming units (CFUs) per lOOul, for example, about 25 million to about 1.5 billion live CFUs per lOOul, about 25 million to about 1 billion live CFUs per lOOul, about 50 million to about 1.5 billion live CFUs per lOOul, about 50 million to about 1 billion live CFUs per lOOul, or about 50 million to about 2 billion live CFUs per lOOul. Additional examples of suitable unit dosages include but are not limited to, a total of 200 billion to 500 billion colony forming units (CFUs) per dose (e.g., 200 billion, 225 billion, 250 billion, 300 billion, 350 billion, 400 billion, 450 billion, or 500 billion CFUs per dose).

[0033] In some embodiments, an effective or therapeutically effective dose of an agent or a composition of the present invention can be in a range of from about 1.0 gm to about 15.0 gm for an adult patient, more preferably between about 2.0 gm and about 10.0 gm of the composition. Effective doses can be administered to a subject at any suitable frequency, e.g., at least once a week, preferably once a day. Pediatric dosages may be in the range of about 15% to 90% of adult dosages.

[0034] In an embodiment, the agent, the composition, or a composition thereof is provided and administered at least once per day. In an embodiment, the agent, the composition, or a composition thereof is provided and administered at least twice per day. In an embodiment, the agent, the composition, or a composition thereof is provided and administered at least thrice per day. In an embodiment, the agent, the composition, or a composition thereof is provided and administered at least four times per day. In an embodiment, the agent, the composition, or a composition thereof is provided and administered for a period of at least at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 3 week, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, or at least 10 weeks. The dosing regimen involving the agents and / or the compositions in accordance with the present invention can be varied to achieve a desired result, such as may be determined empirically for a given individual subject, or by extrapolation from data obtained from administering the composition to a clinical or other test population. InAttorney Docket No.: 45301.601 general, the agent and / or the composition of the invention may be administered in any suitable dose amount that is effective as a health supplement, food supplement, food additive, and / or therapeutic agent to prevent, ameliorate, regulate, cure or otherwise treat pain and osteoarthritis in a subject with a sickle cell disease.

[0035] In an embodiment, the agent, the composition, or a composition thereof is used for pain, cartilage, or bone remodeling in SCD. In an embodiment, the agent, the composition, or a composition thereof is in a dry form. In an embodiment, the agent, the composition, or a composition thereof of the invention are usefully employed as probiotic supplements, food additives, and food supplements.

[0036] In an aspect, disclosed is a method for modulating (for example, preventing, treating, or reducing) pain, osteoarthritis, and other complications in a subject with a sickle cell disease, the method including providing and / or administering the subject the agent, the composition, or a combination thereof as shown and described herein. In an embodiment, the method includes providing and administering the subject the agent, the composition, or a combination thereof. In an embodiment, the method includes providing the subject the agent, the composition, or a combination thereof. In an embodiment, the method includes administering the subject the agent, the composition, or a combination thereof. In an embodiment, the subject is a mammal. In an embodiment, the subject is human. In an embodiment, the method can be used as a combination therapy along with any other suitable method / s for treating or preventing pain and osteoarthritis in a subject with a sickle cell disease.

[0037] In an embodiment, the method includes the agent or the composition including any suitable microbiome, for example, at least one probiotic, its pharmaceutically acceptable salts, its derivatives, or a combination thereof. In an embodiment, the microbiome is a genetically engineered microbiome, its pharmaceutically acceptable salts, its derivatives, or a combination thereof. In an embodiment, the method includes the agent or the composition including at least one probiotic, its pharmaceutically acceptable salts, its derivatives, or a combination thereof. In an embodiment, the method includes the probiotic, which is a commercially available probiotic, for example, VISBIOME probiotic, its pharmaceutically acceptable salts or derivatives, or a combination thereof. In an embodiment, the method includes the agent or the composition including VISBIOME probiotic, its pharmaceutically acceptable salts, its derivatives, or a combination thereof.Attorney Docket No.: 45301.601

[0038] In an embodiment, the method is administered at least once per day. In an embodiment, the method is administered at least twice per day. In an embodiment, the method is administered at least thrice per day. In an embodiment, the method is administered at least four times per day. In an embodiment, the method is administered for a period of at least at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 3 week, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks or indefinitely.

[0039] The present disclosure is illustrated and further described in more detail with reference to the following non-limiting examples. Section headings as used in this section and the entire disclosure herein are merely for organizational purposes and are not intended to be limiting.

[0040] Compounds and materials are described using standard nomenclature. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs.

[0041] The following terms are used to describe the invention of the present disclosure. In instances where a term is not specifically defined herein, that term is given an art-recognized meaning by those of ordinary skill applying that term in context to its use in describing the present disclosure.

[0042] The use of the terms “a” and “an” and “the” and similar referents (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. By way of example, "an element" means one element or more than one element.

[0043] It should also be understood that, in certain methods described herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited unless the context indicates otherwise. Furthermore, the terms first, second, etc., as used herein are not meant to denote any particular ordering, but simply for convenience to denote a plurality of, for example, layers.

[0044] The terms “comprising”, “having”, “including”, and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to”) unless otherwise noted.Attorney Docket No.: 45301.601

[0045] The terms “about” or “approximately,” as used herein, is inclusive of the stated value and means within an acceptable range of deviation for the particular value as determined by one of ordinary skill in the art, considering the measurement in question and the error associated with measurement of the particular quantity (i.e., the limitations of the measurement system). For example, “about” can mean within one or more standard deviations, or within ± 10% or 5% of the stated value. Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The endpoints of all ranges are included within the range and independently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention as used herein.

[0046] The phrase "and / or," as used herein in the specification and in the claims, should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with "and / or" should be construed in the same fashion, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and / or" clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to "A and / or B", when used in conjunction with open-ended language such as "comprising" can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0047] As used herein in the specification and in the claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of’ or "exactly one of," or,Attorney Docket No.: 45301.601 when used in the claims, "consisting of," will refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein shall only be interpreted as indicating exclusive alternatives (i.e. , "one or the other but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of."

[0048] As used herein in the specification and in the claims, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from anyone or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified. Thus, as a nonlimiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently "at least one of A and / or B") can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

[0049] The phrase "one or more," as used herein, means at least one, and thus includes individual components as well as mixtures / combinations of the listed components in any combination.

[0050] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients and / or reaction conditions are to be understood as being modified in all instances by the term "about," meaning within 10% of the indicated number (e.g., "about 10%" means 9%-l 1% and "about 2%" means 1.8%-2.2%).

[0051] All percentages and ratios are calculated by weight unless otherwise indicated. All percentages are calculated based on the total composition unless otherwise indicated. Generally, unless otherwise expressly stated herein, "weight" or "amount" as used herein with respect to the percent amount of an ingredient refers to the amount of the raw material comprising the ingredient, wherein the raw material may be described herein to comprise less than and up to 100% activity of the ingredient. Therefore, weight percent of an active in a composition isAttorney Docket No.: 45301.601 represented as the amount of raw material containing the active that is used and may or may not reflect the final percentage of the active, wherein the final percentage of the active is dependent on the weight percent of active in the raw material.

[0052] All ranges and amounts given herein are intended to include subranges and amounts using any disclosed point as an end point. Thus, a range of "1% to 10%, such as 2% to 8%, such as 3% to 5%," is intended to encompass ranges of "1% to 8%," "1% to 5%," "2% to 10%, " and so on. All numbers, amounts, ranges, etc., are intended to be modified by the term "about," whether or not so expressly stated. Similarly, a range given of "about 1% to 10%" is intended to have the term "about" modifying both the 1% and the 10% endpoints. Further, it is understood that when an amount of a component is given, it is intended to signify the amount of the active material unless otherwise specifically stated.

[0053] As used herein, the term “administering” means the actual physical introduction of a composition into or onto (as appropriate) a subject, a host or cell. Any and all methods of introducing the composition into the subject, host or cell are contemplated according to the invention; the method is not dependent on any particular means of introduction and is not to be so construed. Means of introduction are well-known to those skilled in the art, and also are exemplified herein.

[0054] As used herein, “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0055] As used herein, the term “pharmaceutically acceptable” refers to compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction when administered to a subject, preferably a human subject. Preferably, as used herein, the term “pharmaceutically acceptable” means approved by a regulatory agency of a federal or state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0056] As used herein, the terms “treat,” “treating,” and “treatment” include inhibiting the pathological condition, disorder, or disease, e.g., arresting or reducing the development of the pathological condition, disorder, or disease or its clinical symptoms; or relieving the pathological condition, disorder, or disease, e.g., causing regression of the pathological condition, disorder, or disease or its clinical symptoms. These terms also encompass therapy and cure. Treatment meansAttorney Docket No.: 45301.601 any way the symptoms of a pathological condition, disorder, or disease are ameliorated or otherwise beneficially altered. Preferably, the subject in need of such treatment is a mammal, preferably a human.

[0057] As used herein, the term "effective amount" or “therapeutically effective amount” as used interchangeably herein, refers to the amount of a therapy, which is sufficient to reduce or ameliorate the severity and / or duration of a disorder or one or more symptoms thereof, inhibit or prevent the advancement of a disorder, cause regression of a disorder, inhibit or prevent the recurrence, development, onset or progression of one or more symptoms associated with a disorder, detect a disorder, or enhance or improve the prophylactic or therapeutic effect(s) of another therapy (e.g., prophylactic or therapeutic agent). An effective or therapeutically effective amount can require more than one dose.

[0058] Effective or therapeutically effective amounts may vary depending upon the biological effect desired in the individual, condition to be treated, and / or the specific characteristics of the composition according to the present invention and the individual. In this respect, any suitable dose of the composition can be administered to the patient (e.g., human), according to the type of disease to be treated. Various general considerations taken into account in determining the “effective amount” are known to those of skill in the art and are described, e.g., in Gilman et al., eds., Goodman And Gilman’s: The Pharmacological Bases of Therapeutics, 8th ed., Pergamon Press, 1990; and Remington’s Pharmaceutical Sciences, 17th Ed., Mack Publishing Co., Easton, Pa., 1990, each of which is herein incorporated by reference. The dose of the composition according to the present invention desirably comprises about 0.1 mg per kilogram (kg) of the body weight of the patient (mg / kg) to about 400 mg / kg (e.g., about 0.75 mg / kg, about 5 mg / kg, about 30 mg / kg, about 75 mg / kg, about 100 mg / kg, about 200 mg / kg, or about 300 mg / kg). In another embodiment, the dose of the composition according to the present invention comprises about 0.5 mg / kg to about 300 mg / kg (e.g., about 0.75 mg / kg, about 5 mg / kg, about 50 mg / kg, about 100 mg / kg, or about 200 mg / kg), about 10 mg / kg to about 200 mg / kg (e.g., about 25 mg / kg, about 75 mg / kg, or about 150 mg / kg), or about 50 mg / kg to about 100 mg / kg (e.g., about 60 mg / kg, about 70 mg / kg, or about 90 mg / kg).

[0059] The term “subject” or “patient” is used herein to refer to an animal, such as a mammal, including a primate (such as a human, a non-human primate, e.g., a monkey, and a chimpanzee), a non-primate (such as a cow, a pig, a camel, a llama, a horse, a goat, a rabbit, aAttorney Docket No.: 45301.601 sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, and a whale), a bird (e.g., a duck or a goose), and a shark. In an embodiment, the subject is a human, such as a human being treated or assessed for a disease, disorder or condition, a human at risk for a disease, disorder or condition, a human having a disease, disorder or condition, and / or human being treated for a disease, disorder or condition as described herein. In some embodiments, the subject does not suffer from an ongoing autoimmune disease. In one embodiment, the subject is about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years of age. In another embodiment, the subject is about 5-10, 10-15, 15-20, 20-25, 25- 30, 30-35, 35-40, 40-45, 45-50, 50-55, 55-60, 60-65, 65-70, 70-75, 75-80, 80-85, 85-90, 90-95, 95-100 years of age. Values and ranges intermediate to the above recited ranges are also intended to be part of this invention. In addition, ranges of values using a combination of any of the above-recited values as upper and / or lower limits are intended to be included.

[0060] As used herein, a “symptom” of a disease includes any clinical or laboratory manifestation associated with the disease, and is not limited to what a subject can feel or observe.

[0061] All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention as used herein. Unless defined otherwise, technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art of this disclosure.

[0062] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group.

[0063] The composition according to the present invention may be administered to a patient by various routes. Examples of routes of administration include, but are not limited to, parenteral, e.g., intravenous, intradermal, subcutaneous, oral, intranasal (e.g., inhalation), transdermal (e.g.,Attorney Docket No.: 45301.601 topical), transmucosal, and rectal administration. In an embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer.

[0064] The disclosed compositions may be prepared in various forms, such as capsules, suppositories, tablets, food / drink and the like. Optionally, the disclosed compositions may include various pharmaceutically acceptable excipients, such as microcrystalline cellulose, mannitol, glucose, defatted milk powder, polyvinylpyrrolidone, starch and combinations thereof.

[0065] The therapeutically effective compounds as described herein may, in accordance with the disclosure, be administered in single or divided doses by the oral, parenteral or topical routes. Administration of the active compound may range from continuous (intravenous drip) to several administrations per day (for example, Q.I.D.) and may include administration routes such as oral, topical, parenteral, intramuscular, intravenous, sub-cutaneous, transdermal (which may include a penetration enhancement agent), buccal, sublingual, intranasal, intraocular, intrathecal, vaginal, and suppository administration, among other routes of administration. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Enteric coated oral tablets may be used to enhance bioavailability of the compounds from an oral route of administration. The most effective dosage form will depend upon the pharmacokinetics of the particular agent chosen as well as the type, location and severity of disease, condition or symptom, and the health of the patient. Administration of compounds according to the present disclosure as sprays, mists, or aerosols for intra-nasal, intra-tracheal or pulmonary administration may also be used. The present disclosure therefore also is directed to pharmaceutical compositions comprising an effective amount of compound as described herein or a pharmaceutically acceptable salt thereof, optionally in combination with a pharmaceutically acceptable carrier, additive or excipient. Compounds according to the present disclosure may be administered in immediate release or sustained or controlled release forms. Sustained or controlled release forms are preferably administered orally, but also in suppository and transdermal or other topical forms. Intramuscular injections in liposomal form or in depot formulation may also be used to control or sustain the release of compound at an injection site.Attorney Docket No.: 45301.601

[0001] The compositions as described herein may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered in controlled-release formulations. It should also be understood that a specific dosage and treatment regimen for any particular patient will depend on the judgment of the treating physician as based upon a variety of factors, including the activity and bioavailability of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the severity of the particular disease or condition being treated.

[0066] A patient or subject in need of therapy using a compound according to the methods described herein can be treated by administering to the patient (subject) an effective amount of the compound according to the present disclosure, either alone, or in combination with another known therapeutic agent. In an embodiment, the patient or subject is a mammal such as human.

[0067] In certain aspects, the agent or compound is conveniently administered in any suitable unit dosage form, including but not limited to a dosage form containing less than 1 milligrams (mg), 1 mg to 3000 mg, or 5 mg to 500 mg of active ingredient per unit dosage form. An oral dosage of about 25 mg-250 mg is often convenient.

[0068] The concentration of the agent or compound in the drug composition will depend on absorption, distribution, metabolism, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the physician administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time.

[0069] In accordance with any of the embodiments, the composition according to the present invention can be administered orally to a subject in need thereof. Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the compound dissolved in diluents, such as water, saline, or orange juice and include an additive, such as cyclodextrin (e.g., a-, [3-, or y-cyclodextrin, hydroxypropyl cyclodextrin) or polyethylene glycol (e.g., PEG400); (b) capsules, sachets, tablets, lozenges, and troches, each containing aAttorney Docket No.: 45301.601 predetermined amount of the active ingredient, as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; and (e) suitable emulsions and gels. Liquid formulations may include diluents, such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and cornstarch. Tablet forms can include one or more of lactose, sucrose, mannitol, com starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers. Lozenge forms can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like containing, in addition to the active ingredient, such carriers as are known in the art.

[0070] The dose administered to the mammal, particularly human and other mammals, in accordance with the present invention should be sufficient to affect the desired response. One skilled in the art will recognize that dosage will depend upon a variety of factors, including the age, condition or disease state, predisposition to disease, genetic defect or defects, and body weight of the mammal. The size of the dose will also be determined by the route, timing and frequency of administration as well as the existence, nature, and extent of any adverse sideeffects that might accompany the administration of a particular composition and the desired effect. It will be appreciated by one of skill in the art that various conditions or disease states may require prolonged treatment involving multiple administrations.EXAMPLES

[0071] Three months old female healthy control (Ctrl) and SCD mice were gavaged with VISBIOME probiotic (dissolved in Medium Chain Triclyceride, MCT oil) or vehicle (MCT oil only), 1 billion CFU per dose, twice per week, for 6 weeks.

[0072] VISBIOME probiotic (EXE-346; ExeGi Pharma, Rockville, MD) comprises 4 strains of Lactobacillus (L. casei subsp. paracasei, L. plantarum, L. acidophilus, and L.Attorney Docket No.: 45301.601 delbrueckii subsp. bulgaricus , three strains of Bifidobacterium (B. longum, B. infantis, and B. breve), one strain of Streptococcus thermophilus, and starch.

[0073] Five weeks after treatment had commenced, pain behaviors were measured (Figure 1A-D). Grip force analysis by computerized grip-meter showed that increased deep tissue hyperalgesia in SCD mice was not rescued by VISBIOME probiotic. Von Frey analysis showed that increased mechanical hyperalgesia in SCD mice was rescued by VISBIOME probiotic. Cold and hot plate analysis showed that increased thermal hyperalgesia in SCD was rescued by VISBIOME probiotic. At 6 weeks post treatment, mice were sacrificed. Digital X-ray images of the knee joint showed that the flattened tibia plateau and uneven joint surfaces in the SCD-Veh mice were partially rescued with VISBIOME probiotic (Figure 2A). In vivo DXA analysis showed decreased femoral bone mineral density (BMD) in SCD mice was not rescued by VISBIOME probiotic. However, micro-CT analysis on excised knee joints showed erosion of articular surface in the SCD-Veh group on gross examination of 3D reconstructions micro-CT images, that was partially rescued by VISBIOME probiotic treatment (Figure 2B).Morphometric parameters calculated from 3 -dimensional Micro-CT of femoral epiphyses showed significantly reduced trabecular thickness (Tb.Th) and tissue mineral density (TMD) in femoral subchondral bone of SCD-Veh, that was partially rescued by VISBIOME probiotic (Figure 3). This illustrates that probiotics such as VISBIOME probiotic are beneficial in SCD- related pain and joint pathology.

[0074] For reasons of completeness, various embodiments of the disclosure are set out in the following numbered clauses:

[0075] Clause 1. A method for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising: administering a therapeutically effective amount of a composition comprising Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus, or any combinations thereof to a subject in need thereof under conditions such that pain and / or symptoms of OA are prevented, treated, or reduced in the subject.

[0076] Clause 2. A method for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising: administering a therapeutically effective amount of a composition comprising Lactobacillus casei subsp. paracasei, Lactobacillus plantarum,Attorney Docket No.: 45301.601Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus to a subject in need thereof under conditions such that pain and / or symptoms of OA are prevented, treated, or reduced in the subject.

[0077] Clause 3. The method of clause 1 or 2, wherein the subject has sickle cell disease.

[0078] Clause 4. The method of any one of clauses 1 to 3, wherein the composition is a pharmaceutical composition.

[0079] Clause 5. The method of clause 4, wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers.

[0080] Clause 6. The method of clause 5, wherein the carrier is a starch.

[0081] Clause 7. The method of any one of the preceding clauses, wherein theLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus are lyophilized.

[0082] Clause 8. The method of any one of the preceding clauses, wherein the composition is a powder, capsule, liquid, or paste.

[0083] Clause 9. The method of any one of the preceding clauses, wherein the administering is oral administration.

[0084] Clause 10. The method of any one of the preceding clauses, wherein the composition is provided as a unit dosage.

[0085] Clause 11. The method of clause 10, wherein each unit dosage comprises a total of about 200 billion to about 500 billion colony forming units (CFUs) per dose.

[0086] Clause 12. The method of clause 11, wherein each unit dosage comprises about 225 billion CFUs per dose.

[0087] Clause 13. The method of clause 11, wherein each unit dosage comprises about 450 billion CFUs per dose.

[0088] Clause 14. The method of any one of the preceding clauses, wherein 1 to 3 doses are administered per day.

[0089] Clause 15. The method of any one of the preceding clauses, wherein the administration is performed for a time period of about 1 week to one year.Attorney Docket No.: 45301.601

[0090] Clause 16. The method of any one of the preceding clauses, wherein the administration is performed indefinitely.

[0091] Clause 17. The method of any one of the preceding clauses, wherein the pain is hyperalgesia and / or hypersensitivity to cold.

[0092] Clause 18. The method of any one of the preceding clauses, wherein the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular bone.

[0093] Clause 19. A composition for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus, or any combinations thereof.

[0094] Clause 20. A composition for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus.

[0095] Clause 21. The composition of clause 19 or 20, wherein the composition is a pharmaceutical composition.

[0096] Clause 22. The composition of clause 21, wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers.

[0097] Clause 23. The composition of clause 22, wherein the carrier is a starch.

[0098] Clause 24. The composition of any one of clauses 19 to 23, wherein theLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus are lyophilized.

[0099] Clause 25. The composition of any one of clauses 19 to 24, wherein the composition is a powder, capsule, liquid, or paste.

[0100] Clause 26. The composition of any one of clauses 19 to 25, wherein the composition is provided as a unit dosage.Attorney Docket No.: 45301.601

[0101] Clause 27. The composition of clause 26, wherein each unit dosage comprises a total of about 200 billion to 500 billion colony forming units (CFUs) per dose.

[0102] Clause 28. The composition of clause 27, wherein each unit dosage comprises about 225 billion CFUs per dose.

[0103] Clause 29. The composition of clause 27, wherein each unit dosage comprises abut 450 billion CFUs per dose.

[0104] Clause 30. The method of any one of clauses 19 to 29 wherein the pain is hyperalgesia and / or hypersensitivity to cold.

[0105] Clause 31. The composition of any one of clauses 19 to 30, wherein the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular bone.

[0106] Clause 32. The use of a composition comprisingLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus, or any combinations thereof for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject.

[0107] Clause 33. The use of a composition comprisingLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject.

[0108] Clause 34. The use of clause 32 or 33, wherein the subject has sickle cell disease.

[0109] While the invention has been described with reference to an exemplary embodiment, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the essential scope thereof. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention will include all embodiments falling within the scope of the appended claims. Any combination of theAttorney Docket No.: 45301.601 above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.Incorporation by Reference

[0110] All U.S. and PCT patent publications and U.S. patents mentioned herein are hereby incorporated by reference in their entirety as if each individual patent publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.Other Embodiments

[0111] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.

Claims

Attorney Docket No.: 45301.601CLAIMSWhat is claimed is:

1. A method for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising: administering a therapeutically effective amount of a composition comprising a Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus to a subject in need thereof under conditions such that pain and / or symptoms of OA are prevented, treated, or reduced in the subject.

2. The method of claim 1, wherein the subject has sickle cell disease.

3. The method of claim 1 or 2, wherein the composition is a pharmaceutical composition.

4. The method of claim 3, wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers.

5. The method of claim 4, wherein the carrier is a starch.

6. The method of any one of the preceding claims, wherein theLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus are lyophilized.

7. The method of any one of the preceding claims, wherein the composition is a powder, capsule, liquid, or paste.

8. The method of any one of the preceding claims, wherein the administering is oral administration.Attorney Docket No.: 45301.6019. The method of any one of the preceding claims, wherein the composition is provided as a unit dosage.

10. The method of claim 9, wherein each unit dosage comprises a total of about 200 billion to about 500 billion colony forming units (CFUs) per dose.

11. The method of claim 10, wherein each unit dosage comprises about 225 billion CFUs per dose.

12. The method of claim 10, wherein each unit dosage comprises about 450 billion CFUs per dose.

13. The method of any one of the preceding claims, wherein 1 to 3 doses are administered per day.

14. The method of any one of the preceding claims, wherein the administration is performed for a time period of about 1 week to one year.

15. The method of any one of the preceding claims, wherein the administration is performed indefinitely.

16. The method of any one of the preceding claims, wherein the pain is hyperalgesia and / or hypersensitivity to cold.

17. The method of any one of the preceding claims, wherein the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular bone.

18. A composition for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject, comprising Lactobacillus casei subsp. paracasei, LactobacillusAttorney Docket No.: 45301.601 plantation, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus or any combination thereof.

19. The composition of claim 18, wherein the composition is a pharmaceutical composition.

20. The composition of claim 19, wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers.

21. The composition of claim 20, wherein the carrier is a starch.

22. The composition of any one of claims 18 to 22, wherein theLactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, and Streptococcus thermophilus are lyophilized.

23. The composition of any one of claims 18 to 22, wherein the composition is a powder, capsule, liquid, or paste.

24. The composition of any one of claims 18 to 23, wherein the composition is provided as a unit dosage.

25. The composition of claim 24, wherein each unit dosage comprises a total of about 200 billion to about 500 billion colony forming units (CFUs) per dose.

26. The composition of claim 25, wherein each unit dosage comprises about 225 billion CFUs per dose.

27. The composition of claim 25, wherein each unit dosage comprises about 450 billion CFUs per dose.Attorney Docket No.: 45301.60128. The composition of any one of claims 18 to 27, wherein the pain is hyperalgesia and / or hypersensitivity to cold.

29. The composition of any one of claims 18 to 28, wherein the symptoms of OA are one or more of flattened tibia plateau, uneven joint surfaces, erosion of articular surface, femoral epiphysis structure, or thinning of trabecular bone.

30. The use of a composition comprising Lactobacillus casei subsp. paracasei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus or any combination thereof for preventing, treating, or reducing pain and / or symptoms of osteoarthritis in a subject.

31. The use of claim 30, wherein the subject has sickle cell disease.

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